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无饲养层培养胚胎干细胞在胶原接枝三维纳米纤维支架上促进干性和多能性。

The promotion of stemness and pluripotency following feeder-free culture of embryonic stem cells on collagen-grafted 3-dimensional nanofibrous scaffold.

机构信息

Stem Cell Biology Department, Stem Cell Technology Research Center, Tehran, Iran.

出版信息

Biomaterials. 2011 Oct;32(30):7363-74. doi: 10.1016/j.biomaterials.2011.06.048. Epub 2011 Jul 16.

DOI:10.1016/j.biomaterials.2011.06.048
PMID:21762983
Abstract

The components of extracellular matrix (ECM) may substitute for feeder layers that promote the self-renewal pathways in embryonic stem cells. Surface modification of electrospun nanofibrous scaffolds have been studied to closely resemble natural ECMs and support in vitro and in vivo proliferation, pluripotency and differentiation of stem cells. In this study, we analyzed the maintenance of stemness and pluripotency of the mouse embryonic stem cell (mESC) following feeder-free culture on collagen-grafted polyethersulfone (PES-COL) electrospun nanofibrous scaffold. Our results showed that, the mESCs cultured for seven passages on PES-COL scaffolds had a typical undifferentiated morphology, enhanced proliferation, stable diploid normal karyotype, and continued expression of stemness and pluripotency-associated markers, Oct-4, Nanog, SSEA-1, and Alkaline phosphatase (ALP) in comparison with PES scaffolds and gelatin-coated plate. Moreover, these cells retained their in vitro and in vivo pluripotency. Our results indicated the enhanced infiltration and teratoma formation of mESCs in PES-COL. Collagen-grafted polyethersulfone nanofibrous scaffold has potential for feeder-free culture of pluripotent stem cells because of its 3-dimensional structure and bioactivity which enhance pluripotency, proliferation, differentiation, and infiltration of embryonic stem cells.

摘要

细胞外基质(ECM)的成分可以替代饲养层,促进胚胎干细胞的自我更新途径。已经研究了静电纺纳米纤维支架的表面修饰,以使其类似于天然 ECM,并支持干细胞的体外和体内增殖、多能性和分化。在这项研究中,我们分析了在胶原接枝聚醚砜(PES-COL)静电纺纳米纤维支架上无饲养层培养后,维持小鼠胚胎干细胞(mESC)干性和多能性。我们的结果表明,与 PES 支架和明胶涂覆板相比,在 PES-COL 支架上培养 7 代的 mESCs 具有典型的未分化形态、增强的增殖、稳定的二倍体正常核型,以及持续表达干性和多能性相关标志物 Oct-4、Nanog、SSEA-1 和碱性磷酸酶(ALP)。此外,这些细胞保留了它们的体外和体内多能性。我们的结果表明,mESCs 在 PES-COL 中的浸润和畸胎瘤形成增强。胶原接枝聚醚砜纳米纤维支架具有作为无饲养层培养多能干细胞的潜力,因为其 3 维结构和生物活性增强了胚胎干细胞的干性、增殖、分化和浸润。

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